Archives of Microbiology

, Volume 143, Issue 4, pp 330–336 | Cite as

Aspects of nitrogen fixation in Chlorobium

  • Ghanshyam D. Heda
  • Michael T. Madigan
Original Papers


Four strains of the green sulfur bacterium Chlorobium were studied in respect to nitrogen nutrition and nitrogen fixation. All strains grew on ammonia, N2, or glutamine as sole nitrogen sources; certain strains also grew on other amino acids. Acetylene-reducing activity was detectable in all strains grown on N2 or on amino acids (except for glutamine). In N2 grown Chlorobium thiosulfatophilum strain 8327 1 mM ammonia served to “switch-off” nitrogenase activity, but the effect of ammonia was much less dramatic in glutamate or limiting ammonia grown cells. The glutamine synthetase inhibitor methionine sulfoximine inhibited ammonia “switch-off” in all but one strain. Cell extracts of glutamate grown strain 8327 reduced acetylene and required Mg2+ and dithionite, but not Mn2+, for activity. Partially purified preparations of Rhodospirillum rubrum nitrogenase reductase (iron protein) activating enzyme slightly stimulated acetylene reduction in extracts of strain 8327, but no evidence for an indigenous Chlorobium activating enzyme was obtained. The results suggest that certain Chlorobium strains are fairly versatile in their nitrogen nutrition and that at least in vivo, nitrogenase activity in green bacteria is controlled by ammonia in a fashion similar to that described in nonsulfur purple bacteria and in Chromatium.

Key words

Phototrophic bacteria Green sulfur bacteria Chtorobium Nitrogen fixation Nitrogenase 

Non-common abbreviations


Methionine sulfoximine


3-(N-morpholino) propane sulfonic acid


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  1. Alef K, Kleiner D (1982) Regulatory aspects of inorganic nitrogen metabolism in Rhodospirillaceae. Arch Microbiol 133:239–241Google Scholar
  2. Arp DJ, Zumft WG (1983) Overproduction of nitrogenase by nitrogen-limited cultures of Rhodopseudomonas palustris. J Bacteriol 153:1322–1330Google Scholar
  3. Belousova AA (1968) Effect of some amino acids on the yield of green sulphur bacteria Chlorobium thiosulfatophilum. Microbiology (English translation of Mikrobiologiya) 37:855–860Google Scholar
  4. Biebl H, Pfennig N (1978) Growth yields of green sulfur bacteria in mixed cultures with sulfur and sulfate reducing bacteria. Arch Microbiol 117:9–16Google Scholar
  5. Brock TD (1978) Thermophilic microorganisms and life at high temperatures. Springer, Berlin Heidelberg New YorkGoogle Scholar
  6. Dowling TE, Preston GG, Ludden PW (1982) Heat activation of the Fe protein of nitrogenase from Rhodospirillum rubrum. J Biol Chem 257:13987–13992Google Scholar
  7. Evans MCW, Smith RV (1971) Nitrogen fixation by the green photosynthetic bacterium Chloropseudomonas ethylicum. J Gen Microbiol 65:95–98Google Scholar
  8. Evans MCW, Telfer A, Cammack R, Smith RV (1971) EPR studies of nitrogenase: ATP dependent oxidation of fraction 1 protein by cyanide. FEBS Letters 15:317–319Google Scholar
  9. Falk G, Johansson BoC, Nordlund S (1982) The role of glutamine synthetase in the regulation of nitrogenase activity (switch-off effect) in Rhodospirillum rubrum. Arch Microbiol 132:251–253Google Scholar
  10. Gest H, Favinger JL, Madigan MT, (1985) Exploitation of N2-fixation capacity for enrichment of anoxygenic photosynthetic bacteria in ecological studies. FEMS Microbiol Ecology 31:317–322Google Scholar
  11. Gest H, Kamen MD (1949) Photoproduction of molecular hydrogen by Rhodospirillum rubrum. Science 109:558–559Google Scholar
  12. Gotto JW, Yoch DC (1985) Regulation of nitrogenase activity by covalent modification in Chromatium vinosum. Arch Microbiol 41:40–43Google Scholar
  13. Gray BH, Fowler CF, Nugent NA, Rigopoulous N, Fuller RC (1973) Reevaluation of Chloropseudomonas ethylica strain 2-K. Intl J Syst Bacteriol 23:256–264Google Scholar
  14. Herbert RA, Siefert E, Pfennig N (1978) Nitrogen assimilation in Rhodopseudomonas acidophila. Arch Microbiol 119:1–5Google Scholar
  15. Hillmer P, Gest H (1977) H2 metabolism in the photosynthetic bacterium Rhodopseudomonas capsulata: H2 production by growing cells. J Bacteriol 129:724–731Google Scholar
  16. Jones BL, Monty KJ (1979) Glutamine as a feedback inhibitor of the Rhodopseudomonas sphaeroides nitrogenase system. J Bacteriol 139:1007–1013Google Scholar
  17. Kamen MD, Gest H (1949) Evidence for a nitrogenase system in the photosynthetic bacterium Rhodospirillum rubrum. Science 109:560Google Scholar
  18. Kelly DP (1974) Growth and metabolism of the obligate photolithotroph Chlorobium thiosulfatophilum in the presence of added organic nutrients. Arch Microbiol 100:163–178Google Scholar
  19. Keppen OI, Lebedeva NV, Petukhov SA, Rodionov YuV (1985) The activity of nitrogenase in the green sulfur bacterium Chlorobium limicola forma thiosulfatophilum (In Russian). Mikrobiologiya 54:36–41Google Scholar
  20. Kovacs KL, Bagyinka Cs, Serebriakova LT (1983) Distribution and orientation of hydrogenase in various photosynthetic bacteria. Curr Microbiol 9:215–218Google Scholar
  21. Lindstrom ES, Tove SR, Wilson PW (1950) Nitrogen fixation by the green and purple sulfur bacteria. Science 112:197–198Google Scholar
  22. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275Google Scholar
  23. Ludden PW, Burris RH (1976) Activating factor for the iron protein of nitrogenase from Rhodospirillum rubrum. Science 194:424–426Google Scholar
  24. Ludden PW, Burris RH (1978) Purification and properties of nitrogenase from Rhodospirillum rubrum, and evidence for phosphate, ribose and an adenine-like unit covalently bound to the iron protein. Biochem J 175:251–259Google Scholar
  25. Madigan MT, Cox SS, Stegeman RA (1984) Nitrogen fixation and nitrogenase activities in members of the family Rhodospirillaceae. J Bacteriol 157:73–78Google Scholar
  26. Madigan MT, Gest H (1979) Growth of the photosynthetic bacterium Rhodopseudomonas capsulata chemoautotrophically in darkness with H2 as the energy source. J Bacteriol 137:524–530Google Scholar
  27. Masters RA, Madigan MT (1983) Nitrogen metabolism in the phototrophic bacteria Rhodocyclus purpureus and Rhodospirillum tenue. J Bacyeriol 155:222–227Google Scholar
  28. Pfennig N, Biebl H (1976) Desulfuromonas acetoxidans, gen. nov., and sp. nov., a new anaerobic, sulfur-reducing, acetate-oxidizing bacterium. Arch Microbiol 110:3–12Google Scholar
  29. Pfennig N, Trüper HG (1981) Isolation of members of the families Chromatiaceae and Chlorobiaceae. In: Starr MP, Stolp H, Trüper HG, Balows A, Schlegel HG (eds) The prokaryotes, a handbook on habitats, isolation and identification of bacteria, vol I. Springer, Berlin Heidelberg New York, pp 279–289Google Scholar
  30. Postgate JR (1982) The fundamentals of nitrogen fixation. Cambridge University Press, Cambridge, EnglandGoogle Scholar
  31. Reiderer-Henderson MA, Wilson PW (1970) Nitrogen fixation by sulfate-reducing bacteria. J Gen Microbiol 61:27–31Google Scholar
  32. Saari LL, Triplett EW, Ludden PW (1984) Purification and properties of the activating enzyme for iron protein of nitrogenase from the photosynthetic bacterium Rhodospirillum rubrum. J Biol Chem 259:15502–15508Google Scholar
  33. Simpson FB, Burris RH (1984) A nitrogen pressure of 50 atmospheres does not prevent evolution of hydrogen by nitrogenase. Science 224:1095–1097Google Scholar
  34. Smith RV, Telfer A, Evans MCW (1971) Complementary functioning of nitrogenase components from a blue-green alga and a photosynthetic bacterium. J Bacteriol 107:574–575Google Scholar
  35. Stackebrandt E, Woese CR (1981) The evolution of prokaryotes. In: Carlile MJ, Collins JR, Moseley BEB (eds) Molecular and cellular aspects of microbial evolution. Cambridge University Press, pp 1–31Google Scholar
  36. Stegeman RA, Madigan MT (1985) Nitrogen nutrition and pathway of ammonia assimilation in brown Rhodospirillum species. FEMS Microbiology Letts 26:259–264Google Scholar
  37. Sweet WJ, Burris RH (1981) Inhibition of nitrogenase activity by NH4+ in Rhodospirillum rubrum. J Bacteriol 145:824–831Google Scholar
  38. Wagner BJ, Miović ML, Gibson J (1973) Utilization of amino acids by Chromatium sp. strain D. Arch Mikrobiol 91:255–272Google Scholar
  39. Yoch DC (1979) Manganese, an essential trace element for N2 fixation by Rhodospirillum rubrum and Rhodopseudomonas capsulata: role in nitrogenase regulation. J Bacteriol 140:987–995Google Scholar
  40. Yoch DC (1980) Regulation of nitrogenase A and R concentrations in Rhodopseudomonas capsulata by glutamine synthetase. Biochem J 187:273–276Google Scholar
  41. Zakhvataeva NV, Kondrateva EN (1971) Fixation of molecular nitrogen by photosynthesizing bacteria in relation to presence of light and ATP and character of exogenous substrate. Dokl Akad Nauk SSSR 196:72–74Google Scholar
  42. Zakhvataeva NV, Malofeeva IV, Kondrateva EN (1970) Nitrogen fixation capacity of photosynthesizing bacteria. Microbiology (English translation of Mikrobiologiya) 39:661–666Google Scholar

Copyright information

© Springer-Verlag 1986

Authors and Affiliations

  • Ghanshyam D. Heda
    • 1
  • Michael T. Madigan
    • 1
  1. 1.Department of MicrobiologySouthern Illinois UniversityCarbondaleUSA

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